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The evaluation of the biogenic amines (BAs) profile of different types of craft beers is herein presented. A previously developed and validated analytical method based on ion-pair chromatography coupled with potentiometric detection was used to determine the presence of 10 BAs. Good analytical features were obtained for all amines regarding linearity (R2 values from 0.9873 ± 0.0015 to 0.9973 ± 0.0015), intra- and inter-day precision (RSD lower than 6.9% and 9.7% for beer samples, respectively), and accuracy (recovery between 83.2-108.9%). Detection and quantification limits range from 9.3 to 60.5 and from 31.1 to 202.3 µg L-1, respectively. The validated method was applied to the analysis of four ale beers and one lager craft beer. Ethylamine, spermidine, spermine, and tyramine were detected in all analyzed samples while methylamine and phenylethylamine were not detected. Overall, pale ale beers had a significantly higher total content of BAs than those found in wheat pale and dark samples. A general least square regression model showed a good correlation between the total content of BAs and the brewing process, especially for Plato degree, mashing, and fermentation temperatures. Knowledge about the type of ingredients and manufacturing processes that contribute to higher concentrations of these compounds is crucial to ensuring consumer safety.
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Cerveja , Aminas Biogênicas , Cerveja/análise , Aminas Biogênicas/análise , Cromatografia Líquida de Alta Pressão/métodos , Espermidina/análise , Espermina/análiseRESUMO
A supramolecular atropine sensor was developed, using cucurbit[6]uril as the recognition element. The solid-contact electrode is based on a polymeric membrane incorporating cucurbit[6]uril (CB[6]) as an ionophore, 2-nitrophenyl octyl ether as a solvent mediator, and potassium tetrakis (4-chlorophenyl) borate as an additive. In a MES-NaOH buffer at pH 6, the performance of the atropine sensor is characterized by a slope of (58.7 ± 0.6) mV/dec with a practical detection limit of (6.30 ± 1.62) × 10-7 mol/L and a lower limit of the linear range of (1.52 ± 0.64) × 10-6 mol/L. Selectivity coefficients were determined for different ions and excipients. The obtained results were bolstered by the docking and spectroscopic studies which demonstrated the interaction between atropine and CB[6]. The accuracy of the potentiometric analysis of atropine content in certified reference material was evaluated by the t-Student test. The herein proposed sensor answers the need for reliable methods providing better management of this hospital drug shelf-life while reducing its flush and remediation costs.
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Atropina , Polímeros , Eletrodos , Humanos , Ionóforos , PotenciometriaRESUMO
End-stage renal disease is a growing health problem with increasing prevalence and high health care costs. Patients suffering from end-stage renal disease exhibit higher morbidity and mortality rates compared to the general population. These patients, who are treated using hemodialysis, typically suffer from anemia, inflammation, and oxidative stress. Inadequate dialyzer membrane biocompatibility exacerbates these negative side effects. Modifications of the composition of hemodialysis membranes have improved their biocompatibility and improve the patients' quality of life. Recently, the use of dialyzer membranes coated with bioactive compounds has also been proposed to further ameliorate dialysis-associated problems. Based on a survey of the current literature, application of bioactive membranes decreases the inflammation and oxidative stress of patients treated with hemodialysis.
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Materiais Biocompatíveis , Falência Renal Crônica/terapia , Membranas Artificiais , Diálise Renal/efeitos adversos , Antioxidantes/administração & dosagem , Celulose , Humanos , Inflamação/etiologia , Inflamação/prevenção & controle , Estresse Oxidativo , Proteínas Secretadas Inibidoras de Proteinases , Qualidade de Vida , Ácido Tióctico/administração & dosagem , Vitamina E/administração & dosagemRESUMO
Canine leishmaniosis (CanL) is a major veterinary concern and a public health issue. Serological data are essential for disease management. Several antigens used in serological assays have specificity related problems preventing relevant seropositivity values establishment. Herein we report significant seropositivity level disparity in a study cohort with 384 dogs from eight countries, for antigens traditionally used in CanL - soluble promastigote Leishmania antigens (SPLA) and K39 recombinant protein (rK39): 43·8 and 2·9% for SPLA and rK39, respectively. To better understand the reasons for this disparity, CanL-associated serological response was characterized using, for complement serological evaluation, a ubiquitous antigen - soluble Escherichia coli antigens (SECAs). Using cohorts of CanL dogs and dogs without clinical evidences of CanL from non-endemic regions of Portugal, the serological response of CanL animals followed specific trend of seropositivity rK39 > SPLA > SECA absent in non-diseased animals. Using receiver operating characteristic curve analysis, these characteristic trends were converted in ratios, SPLA/SECA, rK39/SECA and rK39/SPLA, that presented high predictive for discriminating the CanL cohort that was potentiated when applied in a scoring system involving positivity to four out of five predictors (rK39, SPLA, SPLA/SECA, rK39/SECA and rK39/SPLA). In fact, this approach discriminated CanL with similar sensitivity/specificity as reference antigens, diminishing seropositivity in European cohort to 1·8%. Ultimately, non-related antigens like SECA and seropositivity ratios between antigens enable different perspectives into serological data focusing on the search of characteristic serological signatures and not simple absolute serology values contributing to comprehensive serological status characterization.
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Adenosina Trifosfatases/sangue , Antígenos de Bactérias/sangue , Antígenos de Protozoários/sangue , Proteínas de Bactérias/sangue , Doenças do Cão/diagnóstico , Escherichia coli/imunologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/veterinária , Canais de Translocação SEC/sangue , Animais , Doenças do Cão/imunologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/imunologia , Portugal , Proteínas de Protozoários/sangue , Proteínas Recombinantes/sangue , Proteínas SecA , Sensibilidade e EspecificidadeRESUMO
A vortex-assisted liquid-liquid micro-extraction coupled with high-performance liquid chromatography, with UV-vis, is proposed to pre-concentrate methyl methacrylate and to improve separation in biological matrices. The use of 1-octanol as extracting phase, its volume, the need for a dispersant agent, the agitation conditions and the cooling time before phase separation were evaluated. In optimum conditions, enrichment factors of 20 (±0.5) and enrichment recovery of 99% were obtained. The straightforward association of this extraction process with the HPLC method, previously regulated by the International Organization for Standardization, afforded a detection limit of 122 ng/mL and a quantification limit of 370 ng/mL. The within-batch precision, relative standard deviation, was 3% for a sample with 1.49 µg/mL and 4% for a sample with 13.4 µg/mL. The results showed a between batch-precision of 21% for experiments performed on five different days, for a sample with a concentration of 1.10 µg/mL in methyl methacrylate.
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Cromatografia Líquida de Alta Pressão/métodos , Microextração em Fase Líquida/métodos , Metilmetacrilato/análise , Metilmetacrilato/isolamento & purificação , Saliva/química , Humanos , Sensibilidade e EspecificidadeRESUMO
Helicobacter pylori (H. pylori) infection is highly prevalent worldwide, affecting more than 43% of world population. The infection can be transmitted through different routes, like oral-oral, fecal-oral, and gastric-oral. Electrochemical sensors play a crucial role in the early detection of various substances, including biomolecules. In this study, the development of nanobody (Nb)-based immunosensor for the detection of H. pylori antigens in saliva samples was investigated. The D2_Nb was isolated and characterized using Western blot and ELISA and employed in the fabrication of the immunosensor. The sensor was prepared using gold screen-printed electrodes, with the immobilization of Nb achieved through chemical linkage using cysteamine-glutaraldehyde. The surface of the electrode was characterized using EIS, FTIR and SEM. Initially, the Nb-based immunosensor's performance was evaluated through cyclic voltammetry (CV), differential pulse voltammetry (DPV), and square wave voltammetry (SWV). The sensor exhibited excellent linearity with an R2 value of 0.96. However, further assessment with the DPV technique revealed both a low limit of detection (5.9 ng/mL, <1 cfu/mL) and high selectivity when exposed to a mixture of similar antigens. Moreover, the immunosensor demonstrated robust recovery rates (96.2%-103.4%) when spiked into artificial saliva and maintained its functionality when stored at room temperature for 24 days.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Infecções por Helicobacter , Helicobacter pylori , Limite de Detecção , Saliva , Anticorpos de Domínio Único , Saliva/microbiologia , Saliva/química , Técnicas Biossensoriais/instrumentação , Helicobacter pylori/imunologia , Helicobacter pylori/isolamento & purificação , Humanos , Anticorpos de Domínio Único/química , Anticorpos de Domínio Único/imunologia , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/imunologia , Imunoensaio/métodos , Ouro/química , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/análise , Antígenos de Bactérias/isolamento & purificaçãoRESUMO
In this study, a hybrid material, MIL-53(Al)@CBS, was synthesized via the solvothermal method, involving the growth of MIL-53(Al) crystals on cocoa bean shell residues (CBS). Physicochemical characterization techniques, including TGA, BET, FTIR, XRD, and SEM, confirmed successful hybridization. MIL-53(Al)@CBS was employed as an adsorbent for antibiotics (oxytetracycline, tetracycline, chlortetracycline) separation from aqueous solutions. Parameters like pH, adsorbent dose, concentration, time, and temperature were systematically evaluated. FTIR revealed π-π interactions and hydrogen bonds between tetracyclines and the adsorbent. MIL-53(Al)@CBS exhibited adsorption, with removal rates up to 98.92%, 99.04%, and 98.24% for OTC, TC, and CTC, respectively. Kinetics suggested adsorption depends on active site availability, with TC adsorbing fastest. Microscopic models showed adsorption on three distinct active site types with different affinities without competition or adherence to the Langmuir hypothesis. Importantly, MIL-53(Al)@CBS maintained high adsorption capacity even after ten washing cycles, highlighting its potential for water treatment.
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Oxitetraciclina , Poluentes Químicos da Água , Adsorção , Antibacterianos/química , Tetraciclina , Tetraciclinas , Cinética , Poluentes Químicos da Água/químicaRESUMO
Glyphosate (GLY) is the most widely used non-selective broad-spectrum herbicide worldwide under well-reported side effects on the environment and human health. That's why it's necessary to control its presence in the environment. This work describes the development of an affordable, simple, and accurate electrochemical biosensor using a pencil graphite electrode as support, a horseradish peroxidase enzyme immobilized on a polysulfone membrane doped with multi-walled carbon nanotubes. The developed electrochemical sensor was used in the determination of GLY in river and drinking water samples. Cyclic voltammetry and amperometry were used as electrochemical detection techniques for the characterization and analytical application of the developed biosensor. The working mechanism of the biosensor is based on the inhibition of the peroxidase enzyme by GLY. Under optimal experimental conditions, the biosensor showed a linear response in the concentration range of 0.1 to 10 mg L-1. The limits of detection and quantification are 0.025 ± 0.002 and 0.084 ± 0.007 mg L-1, respectively, which covers the maximum residual limit established by the EPA for drinking water (0.7 mg L-1). The proposed biosensor demonstrated high reproducibility, excellent analytical performance, repeatability, and accuracy. The sensor proved to be selective against other pesticides, organic acids, and inorganic salts. Application on real samples showed recovery rates ranging between 98.18 ± 0.11 % and 97.32 ± 0.23 %. The analytical features of the proposed biosensor make it an effective and useful tool for the detection of GLY for environmental analysis.
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Técnicas Biossensoriais , Água Potável , Grafite , Nanotubos de Carbono , Humanos , Grafite/química , Nanotubos de Carbono/química , Reprodutibilidade dos Testes , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Eletrodos , GlifosatoRESUMO
The development of new materials that have a high capacity to remove pollutants in water-based media is becoming increasingly important because of the serious contamination of water and the negative impact on biodiversity and public health. The presence of glyphosate in water, the most widely used herbicide worldwide, has triggered alerts owing to the collateral effects it may cause on human health. The main objective of the present study was to investigate the potential of the hybrid material MIL-53(Al)@RH for the adsorption of glyphosate in aqueous solution. The material was obtained following the methodology of MIL-53(Al) synthesis in the presence of hydrolyzed rice husk assisted by microwave. Batch adsorption experiments were carried out to evaluate the adsorbent dosage, pH0 solution effect, contact time, adsorbate concentration, and temperature effect. The results demonstrated that a maximum adsorption capacity of 296.95 mg g-1, at pH0 4 with a ratio of 0.04 g MIL-53(Al)@RH/50 mL of solution, was achieved in 30 min. The Avrami and pseudo-second order models appropriately described the adsorption kinetics and the equilibrium by Langmuir and Sips models. The enthalpy changes (ΔH°) determined propose an endothermic reaction governed by chemisorption, corroborating the kinetic and equilibrium settings. Hydrogen bonds, π*-π interactions, and complexation between the metal centers of MIL-53(Al) and the anionic groups of glyphosate were postulated to be involved as adsorption mechanisms. Finally, for practical application, MIL-53(Al)@RH was packed in a column for a fixed-bed test which revealed that the hybrid can remove glyphosate with an adsorption capacity of 76.304 mg L-1, utilizing 90% of the bed.
Assuntos
Oryza , Poluentes Químicos da Água , Purificação da Água , Humanos , Água , Poluentes Químicos da Água/química , Oryza/química , Adsorção , Cinética , Concentração de Íons de Hidrogênio , Purificação da Água/métodos , GlifosatoRESUMO
The use of polysulfone (PSU) hemodialysis (HD) membranes modified with bioactive compounds has gained relevance in chronic kidney disease (CKD) management. Compounds based on the 4-oxo-ß-lactam scaffold have outstanding inhibitory ability and selectivity for human neutrophil elastase (HNE). The present work aimed to evaluate the bioactivity and biocompatibility of PSU-based HD membranes doped with HNE inhibitors (HNEIs). For this, two 4-oxo-ß-lactam derivates (D4L-1 and D4L-2) synthesized in house were used, as well as a commercial HNEI (Sivelestat), for comparison purposes. Their HNE inhibition efficacy was evaluated in in vitro and ex vivo (incubations with human plasma) assay conditions. All biomaterials were bioactive and hemocompatible. The inhibitory capacity of the HNEIs and HNEI-PSU membranes in vitro was D4L-1 > D4L-2 > Sivelestat and D4L-2 > Sivelestat > D4L-1, respectively. In ex vivo conditions, both HNEIs and HNEI-PSU materials presented the same relative inhibitory ability (D4L-1 > D4L-2 > Sivelestat). The difference observed between in vitro and ex vivo conditions is most likely due to the inherent lipophilicity/hydrophobicity of each HNEI influencing their affinity and accessibility to HNE when trapped in the membrane. Compared to Sivelestat, both D4L-1 and D4L-2 (and the respective doped membranes) have more potent inhibition capabilities. In conclusion, this work reports the successful development of PSU membranes functionalized with HNEIs.
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An analytical methodology for detecting ammonium ion (NH4 +) in environmental water through potentiometry-ion chromatography (IC) in tandem is presented here. A multielectrode flow cell is implemented as a potentiometric detector after chromatographic separation of cations in the sample. The electrodes are fabricated via miniaturized all-solid-state configuration, using a nonactin-based plasticized polymeric membrane as the sensing element. The overall analytical setup is based on an injection valve, column, traditional conductometric detector, and new potentiometric detector (in that order), permitting the characterization of the analytical performance of the potentiometric detector while validating the results. The limit of detection was found to be ca. 3 × 10-7 M NH4 + concentration after linearization of the potentiometric response, and intra- and interelectrode variations of <10% were observed. Importantly, interference from other cations was suppressed in the tandem potentiometry-IC, and thus, the NH4 + content in fresh- and seawater samples from different locations was successfully analyzed. This analytical technology demonstrated a great potential for the reliable monitoring of NH4 + at micromolar levels, in contrast to the conductivity detector and previously reported NH4 + potentiometric sensors functioning in batch mode or even coupled with IC. Additionally, the suitability of the potentiometric cell for selective multi-ion analysis in the same sample, i.e., Na+, NH4 +, and K+ in water, has been proven.
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Determination of ten biogenic amines in alcoholic beverages by HPLC coupled to a potentiometric detector for food quality control is herein presented. Biogenic amines were separated by ion-pair chromatography on a C18 column using a gradient mobile phase of acetic acid, acetonitrile, and butane-sulfonic acid. Detection was accomplished by a miniaturized amine-selective electrode. The method was validated following ICH and Eurachem guidelines. Linear regression models provided R2 values from 0.9870 ± 0.0019 to 0.9991 ± 0.0014 for tyramine and cadaverine, respectively. Detection and quantification limits depend on the molecular weight of BAs, ranging from 9.3 to 60.5 and from 31.1 to 202.3 µg L-1 for methylamine and spermine, respectively. Repeatability and intermediate precision showed RSD values lower than 5.8 and 8.3%, respectively. Accuracy of assays yielded recovery values from 86.4 to 109.9%. The biogenic amines content in red wine, white wine, and beer samples were 7.54, 5.24, and 4.58 mg L-1, respectively.
Assuntos
Aminas Biogênicas , Vinho , Bebidas Alcoólicas/análise , Cerveja/análise , Aminas Biogênicas/análise , Cromatografia Líquida de Alta Pressão , Controle de Qualidade , Vinho/análiseRESUMO
Canine leishmaniosis (CanL) is a vector-borne disease caused by Leishmania infantum. Infection in dogs can result in a disease with non-specific clinical signs or in a subclinical condition. Infection diagnosis is crucial to guide public health measures considering the zoonotic potential of L. infantum. Serological approaches to detect infection with a reduced antigen panel potentially limit the quality of the information obtained. To evaluate the impact of using distinct antigens in a serological survey, a cohort with 390 dogs from endemic regions in Portugal was subjected to a serological evaluation using ELISA and DAT. Using ELISA, six Leishmania-specific antigens in conjunction with a non-related antigen, Escherichia coli soluble antigens, were evaluated. The global seroprevalence was 10.5% for DAT and 15.4 to 23.1% for ELISA, depending on the antigen for the latter. Still, only 8.2% of the animals were seropositive to all Leishmania-specific antigens. Importantly, a further 31.0% presented antigen-dependent seropositivity. Considering this observation, a serological score system was proposed and validated to address the complex serology results. With this system, the overall dog seropositivity was 26.9%. This work highlights the limitations of single-antigen serological surveys and presents an approach that might contribute to the establishment of CanL-specific serological profiles.
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Drinking water sources are increasingly subject to various types of contamination due to anthropogenic factors and require proper treatment to remove disease-causing agents. Public drinking water systems use different treatment methods to provide safe and quality drinking water to populations. However, they are ineffective in removing contaminants that are considered a danger to the environment and therefore to humans. Several alternative treatment processes have been proposed, such as membrane filtration, as final purification methods. This paper aims to summarize the type of pollutant compounds, filtration processes, and membranes that have been most studied in this area with particular emphasis on how the modification of membranes, either the manufacturing process or the incorporation of nanomaterials, influences their performance.
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Glyphosate (GLY) is the main ingredient in the weed killer Roundup and the most widely used pesticide in the world. Studies of the harmful effects of GLY on human health began to become more wide-ranging after 2015. GLY is listed by the International Agency for Research on Cancer (IARC) as a carcinogenic hazard to humans. Moreover, GLY has the property to complex with transition metals and are stable for long periods, being considered a high-risk element for different matrices, such as environmental (soil and water) and food (usually genetically modified crops). Since that, it was noticed an increment in the development of new analytical methods for its determination in different matrices like food, environmental and biological fluids. Noteworthy, the application of electrochemical techniques for downstream detection sparked interest due to the ability to minimize or eliminate the use of polluting chemicals, using simple and affordable equipment. This work aims to review the contribution of the electroanalytical methods for the determination of GLY in different food and environmental matrices. Parameters such as the electrochemical transduction techniques based on the electrical measurement signals, receptor materials for electrodes preparation, and the detection mechanisms are described in this review. The literature review shows that the electrochemical sensors are powerful detection system that can be improved by their design and by their portability to fulfil the needs of the GLY determination in laboratory benches, or even in situ analysis.
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Produtos Agrícolas , Glicina , Técnicas Eletroquímicas , Glicina/análogos & derivados , Humanos , Plantas Geneticamente Modificadas , GlifosatoRESUMO
The reduction of free radicals by bioactive membranes used for hemodialysis treatment is an important topic due to the constant rise of oxidative stress-associated cardiovascular mortality by hemodialysis patients. Therefore, it is urgent to find an effective solution that helps to solve this problem. Polysulfone membranes enriched with α-lipoic acid, α-tocopherol, and with both components are fabricated by spin coating. The antioxidant properties of these membranes are evaluated in vitro by determining the lipid-peroxidation level and the total antioxidant status of the blood plasma. The biocompatibility is assessed by quantifying the protein adsorption, platelet adhesion, complement activation, and hemolytic effect. All types of membranes show in vitro antioxidant activity and a trend to reduce oxidative stress in vivo; the best results show membranes prepared with a combination of both compounds and prove to be nonhemolytic and hemocompatible. Moreover, the membrane specific separation ability for the main waste products is not affected by antioxidants incorporation.
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Membranas Artificiais , Estresse Oxidativo/efeitos dos fármacos , Polímeros/farmacologia , Diálise Renal , Sulfonas/farmacologia , Ácido Tióctico/farmacologia , alfa-Tocoferol/farmacologia , Adsorção , Adulto , Antioxidantes/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Proteínas do Sistema Complemento/metabolismo , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Humanos , Ativação Plaquetária/efeitos dos fármacos , Porosidade , Ureia/químicaRESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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Canine leishmaniasis is a major veterinary issue and also a public health challenge due to its zoonotic potential. In this context, serological evaluation is essential for Canine leishmaniasis management. Several serological alternatives, such as rapid diagnostic tests, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence antibody test (IFAT), are well established. In fact, the capacity of distinct tests and antigens, evaluated by their sensitivity and specificity, to detect disease is normally considered sufficient for diagnosing Canine leishmaniasis. In this context, we evaluated the seropositivity using 8 different serological tests (ELISA with Leishmania recombinant proteins (rK39, LicTXNPx); soluble promastigote Leishmania antigens (SPLA); commercial ELISA test) in 82 clinically suspect animals from Northern Portugal. The obtained serological data originated 50% of inconclusive serological information with a mixture of seropositive and seronegative results for individual animals. Cut-off independent risk groups were then generated from the serological data to evaluate the clustering of the samples. This analysis originated risk groups that correlated with the most seropositive samples, suggesting that this method might be used, in a cut-off independent manner, to improve conventional serological evaluation. Ultimately, given that no test prioritization exists, the use of any single serological test increases the potential for misdiagnosis, along with all associated risks for the dog as well as public health. The use of a cut-off independent analysis has the potential to improve the predictive values of these tests, enabling a more accurate evaluation of the dog's condition.
Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/sangue , Doenças do Cão/diagnóstico , Leishmaniose Visceral/sangue , Leishmaniose Visceral/veterinária , Animais , Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Análise por Conglomerados , Doenças do Cão/parasitologia , Cães/parasitologia , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/veterinária , Técnica Indireta de Fluorescência para Anticorpo/métodos , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Leishmania infantum , Proteínas de Protozoários/imunologia , Proteínas Recombinantes/imunologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Testes SorológicosRESUMO
Canine leishmaniosis (CanL) prevention in the Mediterranean basin is considered essential to stop human zoonotic visceral leishmaniasis. In this context, vaccination of dogs is expected to have a significant impact in disease control. CaniLeish® (Virbac Animal Health) is one of a few CanL vaccines that are at this moment licensed in Europe. This vaccine contains purified excreted-secreted proteins of Leishmania having several antigens/immunogens with potential to influence serological response. Therefore, it is important to know if CaniLeish vaccination increased the diagnostic challenges associated with conventional serology, limiting the value of some antigens. To address this 20 dogs from a cohort of 35 healthy dogs that were vaccinated, maintained indoor for 1 month and then returned to their natural domiciles for 2 years. After this period, they were re-called to evaluate their clinical/parasitological condition and assess the evolution of seroreactivity against different antigens: soluble promastigote Leishmania antigens (SPLA), recombinant protein Leishmania infantum cytosolic peroxiredoxin, recombinant protein K39 (rK39), recombinant protein K28 and recombinant kinesin degenerated derived repeat using ELISA. Two years after vaccination all vaccinated non-infected animals were seropositive for SPLA. For the other antigens the serological profile was indistinguishable from non-infected animals. Moreover, vaccinated animals presented a characteristic relative serological profile, with higher normalized serological response to SPLA than rK39. This fact enabled to distinguish with sensitivity 92.3% and specificity 95.4%, vaccinated non-infected dogs from infected and non-infected dogs. Ultimately, relative serological profile enabled the detection of healthy vaccinated animals enabling more accurate serological surveys.
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Pencil leads have been increasingly used as electrode material in electrochemical applications. Commonly denominated as pencil graphite electrodes (PGE), they represent a viable alternative to other standard electrodes due to their comparable electrical properties but mainly for their low cost and availability, enabling disposable applications. In order to achieve the best analytical performance literature evidences the type of lead (hardness level) and electrode surface pre-treatment are critical to the envisaged application. The present review describes the use of PGE in biosensing analysis, more specifically those sensors comprising immobilized enzymes but also briefly referring nucleic acids and other biological entities. It lays an emphasis in the immobilization process of the biological entities while focusing in the analytical performance of each biosensor, mainly sensitivity, linear range and limit of detection as comparative criteria. This review also addresses the main characteristics and properties of PGEs as transducer material in the electrochemical field.